miR-22-3p调控TCTN1基因对皮肤鳞状细胞癌细胞增殖和凋亡的机制研究  

Mechanism of miR-22-3p on proliferation and apoptosis of cutaneous squamous cell carcinoma by regulating TCTN1 gene

作　　者：柴华[1] 李上云[1] 宋丹阳[1] 陈晴燕[1] CHAI Hua;LI Shang-Yun;SONG Dan-Yang;CHEN Qing-Yan(Department of Dermatology,Shenyang Seventh People Hospital,Shenyang 110003,China)

机构地区：[1]沈阳市第七人民医院皮肤科,沈阳110003

出　　处：《中国免疫学杂志》2021年第13期1603-1609,共7页Chinese Journal of Immunology

摘　　要：目的:探讨miR-22-3p对皮肤鳞状细胞癌细胞增殖和凋亡的影响及作用机制。方法:qRT-PCR检测正常皮肤组织和皮肤鳞状细胞癌组织中miR-22-3p和TCTN1 mRNA水平,Western blot检测TCTN1蛋白水平。转染miR-22-3p mimics或果,MTT法检测过表达miR-22-3p或沉默TCTN1表达对A431细胞增殖的影响,流式细胞术检测过表达miR-22-3p或沉默TCTN1表达对A431细胞凋亡的影响,Western blot检测过表达miR-22-3p或沉默TCTN1表达对A431细胞CyclinD1、Ki-67、Cleaved caspase-3蛋白表达的影响。生物信息学软件预测显示TCTN1的3′UTR中含有与miR-22-3p互补的核苷酸序列,双荧光素酶报告基因实验验证miR-22-3p与TCTN1的靶向关系。结果:与正常皮肤组织相比,皮肤鳞状细胞癌组织中miR-22-3p水平明显降低(P<0.05),TCTN1 mRNA和蛋白水平明显升高(P<0.05)。过表达miR-22-3p或沉默TCTN1表达后,A431细胞活性、靶向负调控TCTN1表达。过表达TCTN1逆转了过表达miR-22-3p对A431细胞增殖、凋亡及相关蛋白CyclinD1、Ki-67和Cleaved caspase-3表达的影响。结论:过表达miR-22-3p通过靶向负调控TCTN1表达抑制皮肤鳞状细胞癌A431细胞增殖,并促进其凋亡。Objective:To investigate the effects of miR-22-3 p on proliferation and apoptosis of cutaneous squamous cell carcinoma and its mechanism.Methods:qRT-PCR detected levels of miR-22-3 p and TCTN1 mRNA in normal skin tissues and squamous cell carcinoma tissues,and Western blot detected level of TCTN1 protein.After miR-22-3 p mimics or TCTN1 small interfering RNA was transfected into cutaneous squamous cell carcinoma A431 cells,qRT-PCR detected level of miR-22-3 p or Western blot detected level of TCTN1 protein to verify transfection effect.MTT detected effect of overexpressing miR-22-3 p or silencing TCTN1 on proliferation of A431 cells.Flow cytometry detected effect of overexpressing miR-22-3 p or silencing TCTN1 on apoptosis of A431 cells.Western blot detected effect of overexpressing miR-22-3 p or silencing TCTN1 on expression of CyclinD1,Ki-67 and Cleaved caspase-3 protein in A431 cells.Bioinformatics software predicted that 3’UTR of TCTN1 contains a nucleotide sequence complementary to miR-22-3 p,and dual luciferase reporter gene assay verified the targeting relationship between miR-22-3 p and TCTN1.Results:Compared with normal skin tissues,expression level of miR-22-3 p in cutaneous squamous cell carcinoma was significantly decreased(P<0.05),and expression levels of TCTN1 mRNA and protein were significantly increased(P<0.05).After overexpressing miR-22-3 p or silencing TCTN1,A431 cell activity and expression levels of Ki-67 and CyclinD1 protein were decreased(P<0.05),while cell apoptosis rate and expression level of Cleaved caspase-3 protein were increased(P<0.05).miR-22-3 p negatively regulated TCTN1 expression in A431 cells.Overexpressing TCTN1 reversed the effects of overexpressing miR-22-3 p on proliferation,apoptosis and expression of CyclinD1,Ki-67 and Cleaved caspase-3 protein in A431 cells.Conclusion:Overexpressig miR-22-3 p inhibits proliferation of squamous cell carcinoma A431 cells and promotes apoptosis by negatively regulating TCTN1 expression.

关 键 词：miR-22-3p TCTN1 皮肤鳞状细胞癌 增殖 凋亡 

分 类 号：R739.5[医药卫生—肿瘤]